Accurate information on tree species diversity is critical for forest biodiversity, conservation and management, but mapping forest diversity over large and mixed forest areas using satellite remote sensing data remains a challenge because of scale- and ecosystem-dependent relationships between spectral heterogeneity and tree species diversity. In this study, three different diversity indices (Simpson (λ), Shannon (H’), and Pielou (J’)), were tested to characterize forest tree species diversity using individual monthly and multi-temporal Sentinel-1 and -2 images during 2021. The performance of three different machine learning models, Random Forest (RF), Extreme Gradient Boosting (XGB), and Deep Neural Network (DNN) were tested. A collection of 1,020 plot measurements (comprising 47 tree species and 28,122 trees), randomly collected in a mixed broadleaf-conifer forest area in northeast China, was used to train (n = 816) and validate (n = 204) the models. The models dependent on multi-temporal Sentinel-1/2 imagery were found to outperform the models based on individual monthly data, in predicting forest tree species diversity, with average accuracies of 78% for H’, 77% for λ and 77% for J’. The use of DNN performed marginally better than the XGB and RF models, with accuracies of 81% for H’, 80% for λ and 79% for J’, respectively. Finally, a boosted regression model, involving environmental variable predictors and the DNN-based estimated tree species diversity, showed that on average 63 ± 4% of the spatial variations of tree species diversity was explained by environmental variables, including annual temperature (29.30%), followed by soil fertility (27.03%), snow cover (13.63%) and a digital elevation model (12.33%). Our results highlight that an empirical approach based on machine learning and multi-temporal Sentinel-1/2 data can accurately predict forest tree species diversity and we further show the important roles of air temperature and soil fertility in governing the spatial variability of tree species diversity in a mixed broadleaf-conifer forest setting.